Abstract
Monoclonal antibodies have been raised to rat liver cytochromes P-450 b and c, and rabbit liver cytochrome P-450 form 4. A total of six antibodies have been studied. Each antibody reacted strongly both with its homologous antigen and with microsomal fractions selectively enriched with that antigen by treatment of animals with inducing compounds. However, several of the antibodies showed cross-reactivity, either within or between species. A combination of enzyme-linked immunosorbent assay, immunoadsorption, Western blotting and competitive radioimmunoassay revealed that each of the antibodies reacted with a different epitope. Proteolytic digestion of antigen followed by Western blotting of the peptide fragments enabled antibodies, otherwise identical in their reactivity, to be distinguished. It is concluded that complex structural relationships exist amongst the different isoenzymes of cytochrome P-450 and that epitope mapping will help in characterizing both animal and human cytochromes P-450.
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- Atchison M., Adesnik M. A cytochrome P-450 multigene family. Characterization of a gene activated by phenobarbital administration. J Biol Chem. 1983 Sep 25;258(18):11285–11295. [PubMed] [Google Scholar]
- Atlas S. A., Boobis A. R., Felton J. S., Thorgeirsson S. S., Nebert D. W. Ontogenetic expression of polycyclic aromatic compound-inducible monooxygenase activities and forms of cytochrome P-450 in rabbit. Evidence for temporal control and organ specificity of two genetic regulatory systems. J Biol Chem. 1977 Jul 10;252(13):4712–4721. [PubMed] [Google Scholar]
- Boobis A. R., Brodie M. J., Kahn G. C., Fletcher D. R., Saunders J. H., Davies D. S. Monooxygenase activity of human liver in microsomal fractions of needle biopsy specimens. Br J Clin Pharmacol. 1980 Jan;9(1):11–19. doi: 10.1111/j.1365-2125.1980.tb04790.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boobis A. R., McQuade J., Sesardic D., Robson R. T., Hayward C., Lock E. A., Elcombe C. R., Rose M. S., Davies D. S. A monoclonal antibody raised to rat liver cytochrome P-448 (form C) which recognises an epitope common to many other forms of cytochrome P-450. Biochem Pharmacol. 1985 May 15;34(10):1671–1681. doi: 10.1016/0006-2952(85)90633-1. [DOI] [PubMed] [Google Scholar]
- Boobis A. R., Nebert D. W., Felton J. S. Comparison of beta-naphthoflavone and 3-methylcholanthrene as inducers of hepatic cytochrome(s) P-448 and aryl hydrocarbon (benzo[a]pyrene) hydroxylase activity. Mol Pharmacol. 1977 Mar;13(2):259–268. [PubMed] [Google Scholar]
- Boobis A. R., Slade M. B., Stern C., Lewis K. M., Davies D. S. Monoclonal antibodies to rabbit liver cytochrome P-448. Life Sci. 1981 Oct 5;29(14):1443–1448. doi: 10.1016/0024-3205(81)90008-4. [DOI] [PubMed] [Google Scholar]
- Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
- Fujii-Kuriyama Y., Mizukami Y., Kawajiri K., Sogawa K., Muramatsu M. Primary structure of a cytochrome P-450: coding nucleotide sequence of phenobarbital-inducible cytochrome P-450 cDNA from rat liver. Proc Natl Acad Sci U S A. 1982 May;79(9):2793–2797. doi: 10.1073/pnas.79.9.2793. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fujino T., Park S. S., West D., Gelboin H. V. Phenotyping of cytochromes P-450 in human tissues with monoclonal antibodies. Proc Natl Acad Sci U S A. 1982 Jun;79(12):3682–3686. doi: 10.1073/pnas.79.12.3682. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fujita V. S., Black S. D., Tarr G. E., Koop D. R., Coon M. J. On the amino acid sequence of cytochrome P-450 isozyme 4 from rabbit liver microsomes. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4260–4264. doi: 10.1073/pnas.81.14.4260. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guengerich F. P., Dannan G. A., Wright S. T., Martin M. V., Kaminsky L. S. Purification and characterization of liver microsomal cytochromes p-450: electrophoretic, spectral, catalytic, and immunochemical properties and inducibility of eight isozymes isolated from rats treated with phenobarbital or beta-naphthoflavone. Biochemistry. 1982 Nov 9;21(23):6019–6030. doi: 10.1021/bi00266a045. [DOI] [PubMed] [Google Scholar]
- Guengerich F. P., Martin M. V. Purification of cytochrome P-450, NADPH-cytochrome P-450 reductase, and epoxide hydratase from a single preparation of rat liver microsomes. Arch Biochem Biophys. 1980 Dec;205(2):365–379. doi: 10.1016/0003-9861(80)90119-8. [DOI] [PubMed] [Google Scholar]
- Haniu M., Ryan D. E., Levin W., Shively J. E. Structures of cysteine-containing peptides in isosafrole-inducible rat hepatic microsomal cytochrome P-450d: sequence homology with 3-methylcholanthrene-induced cytochrome P-450c. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4298–4301. doi: 10.1073/pnas.81.14.4298. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Ryan D. E., Iida S., Wood A. W., Thomas P. E., Lieber C. S., Levin W. Characterization of three highly purified cytochromes P-450 from hepatic microsomes of adult male rats. J Biol Chem. 1984 Jan 25;259(2):1239–1250. [PubMed] [Google Scholar]
- Ryan D. E., Ramanathan L., Iida S., Thomas P. E., Haniu M., Shively J. E., Lieber C. S., Levin W. Characterization of a major form of rat hepatic microsomal cytochrome P-450 induced by isoniazid. J Biol Chem. 1985 May 25;260(10):6385–6393. [PubMed] [Google Scholar]
- Ryan D. E., Thomas P. E., Levin W. Hepatic microsomal cytochrome P-450 from rats treated with isosafrole. Purification and characterization of four enzymic forms. J Biol Chem. 1980 Aug 25;255(16):7941–7955. [PubMed] [Google Scholar]
- Ryan D. E., Thomas P. E., Reik L. M., Levin W. Purification, characterization and regulation of five rat hepatic microsomal cytochrome P-450 isozymes. Xenobiotica. 1982 Nov;12(11):727–744. doi: 10.3109/00498258209038947. [DOI] [PubMed] [Google Scholar]
- Sogawa K., Gotoh O., Kawajiri K., Fujii-Kuriyama Y. Distinct organization of methylcholanthrene- and phenobarbital-inducible cytochrome P-450 genes in the rat. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5066–5070. doi: 10.1073/pnas.81.16.5066. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Song B. J., Fujino T., Park S. S., Friedman F. K., Gelboin H. V. Monoclonal antibody-directed radioimmunoassay of specific cytochromes P-450. J Biol Chem. 1984 Feb 10;259(3):1394–1397. [PubMed] [Google Scholar]
- Song B. J., Gelboin H. V., Park S. S., Friedman F. K. Epitope-relatedness and phenotyping of hepatic cytochromes P-450 with monoclonal antibodies. Biochem J. 1985 Nov 1;231(3):671–676. doi: 10.1042/bj2310671. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas P. E., Reik L. M., Ryan D. E., Levin W. Induction of two immunochemically related rat liver cytochrome P-450 isozymes, cytochromes P-450c and P-450d, by structurally diverse xenobiotics. J Biol Chem. 1983 Apr 10;258(7):4590–4598. [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yuan P. M., Ryan D. E., Levin W., Shively J. E. Identification and localization of amino acid substitutions between two phenobarbital-inducible rat hepatic microsomal cytochromes P-450 by micro sequence analyses. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1169–1173. doi: 10.1073/pnas.80.5.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]



